Rapid and sensitive UHPLC-DAD method for simultaneous determination of sofosbuvir and ledipasvir in human serum

Today, the direct-acting antiviral agents (DAAs) such as sofosbuvir (SOF) and ledipasvir (LED) are widely used to treat the hepatitis virus infection. The aim of this study was to develop a rapid, simple and valid method for simultaneous determination of SOF and LED in human plasma for bioavailability and pharmacokinetic studies. Chromatographic analysis was performed on the C-18 column (Blue Orchid, 1.8 mu m, 50 x 2 mm) using 0.1 % formic acid in water (pH 2.6) and acetonitrile (60:40; v/v) as mobile phase at a flow rate of 0.5 mL/min. The UV detector was set at 328 nm and 260 nm for analysis of SOF and LED, respectively. To 400 mu L of plasma, 100 mu L of clonazepam as the internal standard (I.S, 7 mu g/mL) was added and the mixture subjected to liquid-liquid extraction using 1000 mu L diethyl ether. The calibration curves were linear with coefficients of variation less than 8% for all analyses. The limit of quantification (LOQ) was 20 and 5 ng/mL for SOF and LED, respectively. The results of inter-day and intra-day precision showed good reproducibility and the total analysis time was 1.2 min. This method successfully applied for determination SOF and LED in four healthy volunteers. (C) 2020 Published by Elsevier B.V.

Automated summary

This study aimed to develop a rapid, simple, and accurate method for simultaneous determination of SOF and LED in human plasma for bioavailability and pharmacokinetic studies. The method showed good precision, reproducibility, and was successfully applied to healthy volunteers with a short total analysis time.